Effect of the water-soluble cholesterol-methyl-β-cyclodextrin complex on the development of pulmonary edema with an analysis of the osmotic resistance of erythrocytes in rats

Introduction. Methyl-β-cyclodextrin is a potent acceptor of cellular membrane cholesterol and, at the same time, used as a solubilizer, which makes it a potential target for targeted delivery of hydrophobic compounds.Aim. To assess the effect of administration of methyl-β-cyclodextrin containing cholesterol on the degree of lung hydration and osmotic resistance of erythrocytes against the background of pulmonary edema in adult rats caused by the administration of mezaton.Materials and methods. We investigated the degree of hydration of the lungs by the value of the pulmonary coefficient and their dry residue. Osmotic resistance of erythrocytes were assessed by the intensity of their hemolysis in a series of hypotonic sodium chloride solutions in adult male Wistar rats (220±40 g) of the control group (intact, n=10) and two experimental groups with pulmonary edema caused by the administration of mezaton (n=10) and with the subsequent introduction of a water-soluble complex cholesterol-methyl-β-cyclodextrin against the background of the development of pulmonary edema (n=10). We used additional criteria for assessing the osmotic resistance of erythrocytes: minimum resistance – the concentration of sodium chloride solution, at which the first “weak” erythrocytes begin to hemolyze; maximum resistance is the concentration of sodium chloride solution at which all or almost all erythrocytes hemolyzed.Results. The introduction of mezaton led to the development of pronounced hemolysis and hydration of the lungs with the development of their edema and a narrowing of the range of minimum and maximum osmotic resistance of erythrocytes. The introduction of a complex of cholesterol with methyl-β-cyclodextrin after edemogenic exposure led to a decrease in the severity of pulmonary edema, determined by a decrease in the pulmonary coefficient and an increase in the dry residue, and to an increase in the osmotic resistance of erythrocytes with an expansion of the range of their minimum and maximum osmotic resistance.Conclusion. The introduction of a water-soluble complex of cholesterol with methyl-β-cyclodextrin against the background of the development of hemodynamic pulmonary edema significantly reduced the degree of hydration of the lungs, which combined with an increase in the osmotic resistance of erythrocytes.

Influence of cardiopulmonary bypass on the erythrocyte membranes and the method of its protection

The damage to erythrocytes during cardiopulmonary bypass (CPB) remains a recent problem. The aim of this research was to study the effect of fructose-1,6-diphosphate on the state of the erythrocyte membrane during CPB and the level of phosphorus in blood as a marker of the energy potential in the cell. Patients were divided into two groups. The control group 1 (Gr 1) consisted of 75 individuals. The group 2 (Gr 2) included patients to whom fructose-1,6-diphosphate (FDP) was administrated according to the developed scheme as follows 10 g of the drug was diluted in 50 ml of a solvent, 5 g of the drug was injected intravenously with the use of perfusor immediately before initiation of CPB at a rate of 10 ml/min and 5 g at the 30th minute of CPB (before the stage of warming) the same way. When comparing two groups the best results in hemolysis (p<0.01), mechanical (p<0.01). osmotic resistance of erythrocytes (p<0.01), the time of acid hemolysis (p<0.01) and the permeability of the erythrocyte membrane in postperfusion period were in Gr 2. Вefore cardiac surgery hypophosphatemia was detected in 18% out of 150 and in 32% out of 150 patients – a lower limit of normal phosphorus content in the blood. After CPB in Gr 1 phosphorus content in blood was 0.85±0.32 mmol/l and hypophosphatemia was in 53% out of 75 patients. This indicates a pronounced energy deficit in this group. In Gr 2 phosphorus level was 1.7±0.31 mmol/l and there was no hypophosphatemia. As a result, FDP as an endogenous high-energy intermediate metabolite of the glycolytic pathway leads to resistance to hemolysis, protects the erythrocyte membrane from damage and increases the energy potential of the cell during CPB.

Comparison of Osmotic Resistance, Shape and Transmembrane Potential of Erythrocytes Collected from Healthy and Fed with High Fat-Carbohydrates Diet (HF-CD) Pigs—Protective Effect of Cistus incanus L. Extracts

Long-term high fat-carbohydrates diet (HF-CD) contributes to the formation of irreversible changes in the organism that lead to the emergence of civilization diseases. In this study, the impact of three-month high-fat diet on the physical properties of erythrocytes (RBCs) was studied. Furthermore, the biological activity of Cistus incanus L. extracts, plant known with high pro-health potential, in relation to normal and HF-CD RBCs, was determined. Obtained results have shown that, applied HF-CD modified shape, membrane potential and osmotic resistance of erythrocytes causing changes in membrane lipid composition and the distribution of lipids. The impact of HF-CD on physical properties of RBCs along with atherosclerotic lesions of the artery was visible, despite the lack of statistically significant changes in blood morphology and plasma lipid profile. This suggests that erythrocytes may be good markers of obesity-related diseases. The studies of biological activity of Cistus incanus L. extracts have demonstrated that they may ameliorate the effect of HF-CD on erythrocytes through the membrane-modifying and antioxidant activity.

MEMBRANE-RECEPTOR COMPLEX DYSFUNCTION AS A NEW TARGET FOR THE NEUROTROPHIC THERAPY OF ISCHEMIC STROKE

The aim of the study was the β-adrenergic activity of peripheral blood erythrocyte membranes in the acute period of ischemic stroke (IS). Peripheral blood erythrocytes are the most informative biological tissue for studying many pathogenetic mechanisms, and the work of their membrane-receptor complex (MRC) can very well reflect the functional state of the whole organism, one of the most important criteria for assessing is the adrenergic activity of cytoplasmic membranes in the pathogenesis of IS. This study solves the problem of the finding of the features of changes in the adrenoceptors system in the acute period of IS in the dynamics of the treatment of humans cryopreserved cord blood serum (CCBS). The main scientific results: the obtained results indicate a reduced ability of adrenoceptors (ARs) on erythrocyte membranes to bind blockers due to the desensitisation effect, which is observed from the 1st day of the disease. In the onset of IS, there is a significant increase in the values of β-adrenergic activity of membranes (β-ARM) of erythrocytes in 2.4 times compared with the control. Maximum levels of the indicator (42.43±3.64 CU) are observed in patients with initially severe disease, which indicates significant stress of the sympathoadrenal system (SAS) in these patients. On the 10th day of the disease, there was a decrease in β-ARM relative to the values obtained during hospitalisation. Still, comparing the value of the indicator on the 10th day of IS in both groups, there was an apparent, more significant decrease in β-ARM in the 2nd group of patients, which additionally received CCBS. The use of CCBS contributes to a more pronounced degree of recovery in neurological functions (6.9%) and faster stabilisation of the structure of MRCs in erythrocytes in the form of reducing the values of β-ARM to 16.61±2.86 CU (p<0.05). The area of practical use of the research results: the results of the study can be used to diagnose and treat of the IS. An innovative technological product: A comprehensive approach to assess the β-ARM of peripheral blood erythrocytes in patients with IS by determining the degree of changes in osmotic resistance of erythrocytes (ORE) under the action of adrenergic drugs (AD) is informative and can be used to diagnose of IS. The use of neurotrophic factors is a potentially new method of treating this pathology. Scope of application of the innovative technological product: in the clinical practice of neurological departments for the treatment of patients with strokes

ОСОБЛИВОСТІ ОСМОТИЧНОЇ РЕЗИСТЕНТНОСТІ ЕРИТРОЦИТІВ ТА ВМІСТУ ГЕМОГЛОБІНУ У КРОВІ РИБ ЗА ДІЇ ФЕРУМУ (III)

The osmotic resistance of erythrocytes and the content of hemoglobin in the blood of carp (Cyprinus carpio L.) and pike (Esox lucius L.) under the action of Fe3+ ions at concentrations of 0.2 and 0.5 mg/dm3, were under study which corresponded to 2 and 5 maximum permissible concentration (MPC). The research demonstrated that increased concentrations of Fe3+ ions lead to changes in the osmotic resistance of erythrocytes and the content of hemoglobin in the blood of carp and pike. The findings suggest that high concentrations of Iron (5 MPC) in carp lead to a decrease in the resistance of erythrocyte membranes, and in pike it led to increased osmotic resistance of erythrocytes, which is apparently due to different mechanisms of adaptation of fish erythrocyte membranes to adverse aquatic environment. Species-specificity is also characterized by changes in the content of hemoglobin in the blood of fish species which were researched. An increased amount of pigment in the blood of carp and a decreased amount of hemoglobin in pike under the action of 5 MPC of metal ions was established. The obtained results can be used to assess the physiological condition of fish and water quality under conditions of contamination of watercourses with metals.

Beta-adrenergic reactivity of erythrocytes and the progression of heart failure in patients after myocardial infarction

Aim. To identify the associations between beta-adrenergic reactivity of erythrocytes and the progression of heart failure (HF) in patients after myocardial infarction (MI).Material and methods. The study included 50 patients with HF and history of MI 6 months ago. To determine the level of sympathoadrenal system activity, we analyzed beta-adrenergic reactivity by changing the osmotic resistance of erythrocytes by use of adrenoceptor blocking agent.Results. The frequency of HF progression after index MI was 26% (n=13). All patients were divided into 2 groups depending on the presence/absence of HF progression in the postinfarction period. When determining beta-adrenergic reactivity, it was found that patients with HF progression compared with patients without it had the higher level of beta-adrenergic reactivity of membrane (β-ARM) of erythrocytes: 58,8 (50,9; 78,0) CU and 46,8 (38,0; 66,3) CU, p=0,025). A ROC analysis made it possible to establish the β-ARM level ≥49,53 CU a cut-off point, which can be considered as a marker of HF progression in patients after MI (sensitivity 92,3%, specificity 62,2%). This level of β-ARM is associated with a more than five-fold increase of HF progression risk in patients after MI (OR 5,48; 95% CI 1,28-23,37; p=0,024).Conclusion. In patients with HF and MI history, there is a decrease in the adrenergic reactivity of erythrocyte cell membrane, which is reflected by an increase of β-ARM above normal range of 20 CU. At the same time, β-ARM in patients with HF progression compared with patients without it is significantly increased. Established cut-off point of β-ARM (≥49,53 CU) allows predicting the HF progression with high sensitivity and specificity.

BIOPHYSICAL MARKERS OF ISCHEMIC STROKE

An important and influential factor that directly affects the severity of ischemic stroke (IS) and determines its outcome is the functional state of the membrane-receptor complex (MRC) of cells. One of the most important criteria for assessing this parameter is the β‑adrenergic activity of cytoplasmic membranes (β‑ARM), which plays a leading role in the pathogenesis of IS. The article presents the results of a comprehensive study of the peculiarities of changes in the structural and functional characteristics of brain tissue and β‑adrenoceptors in the acute period of IS to optimize diagnosis. Measurement of changes in the osmotic resistance of erythrocytes (ORE) under the action of β‑blockers was determined by photoelectron colorimetry. The work was based on the materials of a comprehensive examination of 350 patients with the new-onset IS on the 1st, 10th and 21st day of the disease. The severity of the condition and the degree of neurological deficit were objectified using the stroke scale of the National Institutes of Health (NIHSS) with a score in the first hours of the disease, in the dynamics of treatment on the 10th and 21st day. All patients were divided into 2 clinical groups: 1st (n = 183) – patients in moderate severity condition (mean score on the NIHSS scale 11.74 ± 0.33); 2nd (n = 167) – patients in severe condition (mean score on the NIHSS scale 24.06 ± 0.29). As a result of the study, the indicators of β‑ARM of the control group were within normal limits (15.3 ± 4.4 SU), which corresponds to normal β‑ARM. In patients with moderate IS, the indicator exceeded the control values by 1.97 times, which is typical for the average degree of β‑ARM (21–40 SU), and in severe – 2.8 times and was characterized by a low degree of β‑ARM (> 41 SU). An unfavorable sign for the prognosis of the acute period of IS was represented by a further increase in the levels of β‑ARM in the dynamics, which was observed in clinically severe patients. Keywords adrenergic activity, ischemia, erythrocyte, receptor.